Cantuti-Castelvetri Ludovico, Maravilla Erick, Marshall Michael, Tamayo Tammy, D'auria Ludovic, Monge John, Jeffries James, Sural-Fehr Tuba, Lopez-Rosas Aurora, Li Guannan, Garcia Kelly, van Breemen Richard, Vite Charles, Garcia Jesus, Bongarzone Ernesto R
Departments of Anatomy and Cell Biology.
Departments of Anatomy and Cell Biology, Medical Scientist Training Program, University of Illinois at Chicago, Chicago, Illinois 60612, and.
J Neurosci. 2015 Jan 28;35(4):1606-16. doi: 10.1523/JNEUROSCI.2431-14.2015.
The atrophy of skeletal muscles in patients with Krabbe disease is a major debilitating manifestation that worsens their quality of life and limits the clinical efficacy of current therapies. The pathogenic mechanism triggering muscle wasting is unknown. This study examined structural, functional, and metabolic changes conducive to muscle degeneration in Krabbe disease using the murine (twitcher mouse) and canine [globoid cell leukodystrophy (GLD) dog] models. Muscle degeneration, denervation, neuromuscular [neuromuscular junction (NMJ)] abnormalities, and axonal death were investigated using the reporter transgenic twitcher-Thy1.1-yellow fluorescent protein mouse. We found that mutant muscles had significant numbers of smaller-sized muscle fibers, without signs of regeneration. Muscle growth was slow and weak in twitcher mice, with decreased maximum force. The NMJ had significant levels of activated caspase-3 but limited denervation. Mutant NMJ showed reduced surface areas and lower volumes of presynaptic terminals, with depressed nerve control, increased miniature endplate potential (MEPP) amplitude, decreased MEPP frequency, and increased rise and decay rate constants. Twitcher and GLD dog muscles had significant capacity to store psychosine, the neurotoxin that accumulates in Krabbe disease. Mechanistically, muscle defects involved the inactivation of the Akt pathway and activation of the proteasome pathway. Our work indicates that muscular dysfunction in Krabbe disease is compounded by a pathogenic mechanism involving at least the failure of NMJ function, activation of proteosome degradation, and a reduction of the Akt pathway. Akt, which is key for muscle function, may constitute a novel target to complement in therapies for Krabbe disease.
克拉伯病患者骨骼肌萎缩是一种主要的致残表现,会恶化他们的生活质量并限制当前疗法的临床疗效。引发肌肉萎缩的致病机制尚不清楚。本研究使用小鼠(震颤小鼠)和犬类[球状细胞脑白质营养不良(GLD)犬]模型,研究了克拉伯病中导致肌肉退化的结构、功能和代谢变化。使用报告基因转基因震颤-Thy1.1-黄色荧光蛋白小鼠研究了肌肉退化、去神经支配、神经肌肉[神经肌肉接头(NMJ)]异常和轴突死亡情况。我们发现突变肌肉中有大量尺寸较小的肌纤维,且无再生迹象。震颤小鼠的肌肉生长缓慢且微弱,最大力量降低。NMJ中有显著水平的活化半胱天冬酶-3,但去神经支配有限。突变的NMJ显示突触前终末的表面积和体积减小,神经控制减弱,微小终板电位(MEPP)幅度增加,MEPP频率降低,上升和衰减速率常数增加。震颤小鼠和GLD犬的肌肉有显著的能力储存psychosine,这种神经毒素会在克拉伯病中蓄积。从机制上讲,肌肉缺陷涉及Akt通路的失活和蛋白酶体通路的激活。我们的研究表明,克拉伯病中的肌肉功能障碍因一种致病机制而加剧,该机制至少涉及NMJ功能的衰竭、蛋白酶体降解激活和Akt通路的减少。对肌肉功能至关重要的Akt可能构成克拉伯病治疗中补充治疗的新靶点。